DETERMINATION OF OIL CONTENT IN RAPESEEDS USING TWO

METHODS – SOXHLET EXTRACTION AND PULSED NUCLEAR

MAGNETIC RESONANCE SPECTROMETRY

Anna Januszewska, Ewa Maria Siedlecka, Piotr Glamowski,

Izabela Tomasiak
University of Gdansk, Department of Environmental Engineering, Faculty of Chemistry,
ul. Sobieskiego 18, 80–952 Gdansk 6, Poland tel. (0 58) 53 25 437

Introduction
After cereals, oil crops are the second most important source of edible calories for
human societies. They are also sources of many industrial products, and have the potential
to provide raw materials for many more. The four major global traded oil crops are
soybean, oil palm, rapeseed and sunflower, respectively. Together, these four crops
account for 72% of worldwide vegetable-oil production [1]. Therefore an accurate and fast
determination of oil content is important to breeders, growers and buyers.
The traditional method of oil determination in oilseeds is based on solvent
extraction (Soxhlet). This method is laborious, time consuming and uses large volumes of
organic solvents, moreover results depends on operator attention. Currently, the alternative
techniques to solvent extraction are supercritical fluid extraction [2], NIR spectroscopy [3]
and pulsed nuclear magnetic resonance spectrometry [4].
In this article the Soxhlet method was compared with the pulsed nuclear magnetic
resonance spectrometry (pulsed NMR).

Materials and methods

Samples of rapeseeds were taken in accordance with current international methods
of sampling described in ISO standards: PN-EN ISO 542:1997 [5] and PN-EN ISO
664:1997 [6]. In the first instance increment samples ware taken from tracks. The number
of sampling points, where the increment samples are taken, was defined according to lot
size. The sampling points should have been uniformly distributed throughout the lot
volume, according to the principles described in PN-EN ISO 542:1997 (Fig. 1) [5].

Lot size: > 15 tonnes

Number of incremental samples:
5 sampling points

Lot size: 15 – 30 tonnes

Number of incremental samples:
8 sampling points

Lot size: 30 – 50 tonnes

Number of incremental samples:
11 sampling points

Fig. 1. The number of sampling points [5].

 The increment samples were combined and mixed thoroughly to form bulk samples,
which were used to create laboratory samples. Part of laboratory samples were collected
and combined together. The final analytical samples were selected from this mixture [6].
In this paper the analytical samples were marked 1, 2, 3, 4, 5, 6. The total oil
content in rapeseeds was determined in the test samples (5 g), which were obtained from
representative and homogenous analytical samples. Four test samples were taken from
each analytical samples.
The rapeseeds used in the study were acquired from different growers in Poland.

Soxhlet extraction. The solvent extraction was conducted according to official methods
PN-EN ISO 659:1999 [7] and PN-73/R-66164 [8] with petroleum ether (boiling point
40-60ºC) as the extraction solvent. The oil was extracted from 5 g samples in Soxhlet
apparatus. These samples were prepared by drying rapeseed at 103 ±2ºC. The water
contents in all seeds should have been less than 10 % (m/m). For each test sample it was
necessary to carry out three extraction cycles (4 + 2 + 2 hours) to achieve complete oil
recovery from seeds. The oil extraction was repeated for each test sample three times to
ensure full oil recovery. After each extraction cycle seeds were grounded carefully in the
mortar.
The oil percentage of the collected oil in the seeds was determined gravimetrically
and expressed as a weight percent relative to initial weight of the raw oilseeds.

Pulsed nuclear magnetic resonance spectrometry. The total oil content was analyzed
directly on Bruker minispec Pulsed NMR analyzer (operating at 10 MHz) in accordance
with international standard PN-EN ISO 10565:1999 [9]. Five grams of rapeseed were used
for each test. Oilseeds shouldn’t have been cleaned and prepared in any way. The water
contents of all seeds should have been less than 10 % (m/m).
The calibration prior to each series of measurements was checked using three
verification samples of known oil contents [9].
Four analysis were carried out on test samples taken from the same homogenous
analytical sample.

Statistical analysis. In this study standard deviation (SR) was estimated by range (R) and
coefficient kn according to the following formula [10]:

SR = kn · R

The range is the largest value minus the smallest value in a data set. This formula can be
used when the number of determinations (n) is less than 7 [10].
All extraction and analyses were carried out four times. In this paper kn = 0,4857
where n = 4 [10].

Results and Discussion

The oil content in rapeseed was determined using two methods Soxhlet extraction
and pulsed NMR and results are listed in Table 1.
The NMR results are not comparable with Soxhlet solvent extraction data. The
mean values of oil content determined by this method (~ 42,5 %) were lower then the the
Soxhlet extraction (~ 45,7 %).
Table 1. Determination of oil content in rapeseed using two methods – Soxhlet extraction
and pulsed nuclear magnetic resonance spectrometry.

Number Soxhlet method pulsed NMR method

of
analytical
C CM SDR RDS C CM SDR RDS
samples

45,69 35,90
46,21 39,40
1 46,04 0,25 0,54 41,60 6,90 16,60
46,18 41,00
46,09 50,00
45,66 48,80
46,07 43,00 41,50 5,20 12,50
2 45,95 0,19 0,41
46,06 35,90
46,03 38,00
44,59 31,10
44,59 52,60 43,50 10,40 23,90
3 44,59 0,01 0,02
44,60 40,20
44,59 50,20
45,80 39,40
45,78 40,00 38,00 2,30 6,00
4 45,79 0,01 0,02
45,79 37,00
45,80 35,40
46,69 48,00
45,88 44,90 46,90 2,10 4,40
5 46,13 0,39 0,85
45,90 49,30
46,08 45,40
45,20 42,30
46,00 50,10 43,70 6,10 13,90
6 45,47 0,48 0,01
45,01 44,30
45,70 37,60

C – the oil content in test samples, which were taken from the same homogenous
analytical sample [% of dry mass],
CM – mean value of the oil content in test samples [% of dry mass],
SDR – Standard deviation [10],
RDS – Relative standard deviation [%].

The Soxhlet system is repeatable technique for determination of oil content in

rapeseed. The values of standard deviation varied from 0,01 to 0,48. For comparison, SD
and RSD values obtained for NMR analysis are higher and range between 2,10-10,40 and
4,40-23,90 %, respectively. The Soxhlet method has excellent precission with less than 1%
relative standard deviation.
In the case of NMR, data distribution was very high and some statistical errors
occured.
The calibrate procedure cannot be ignored when using this methods. Pulsary NMR is
difficult to calibrate. The calibration curve shall be obtained by measuring the signal from
the three calibration samples with known oil content (Procedure A) [7] or the one standard
of known oil content (Procedure B) [7]. In this study the working calibration curve was
obtained according to Procedure A. The series of standards was prepared in a concentration
range near to the expected unknown concetration (test sample). All samples were measured
in the working range of calibration curve. The correlation coefficient for this linear
regression is high (0,98). It means that the calibration model works properly. There could
be other couses that could affect final results (sample presentation, water content, different
nature of calibration standards then samples, slope of the curve).
In conclusion, the proposed pulsary NMR method is relativelly simple, convenient,
rapid and non-destructive, without sample preparation. This technique offers considerable
savings of solvent disposal costs and analysis time (time of measurments is not more then
2 min.) also eliminates the exposure of laboratory staff to toxic and flammable solvents.
The results in Table 1 suggest that the pulsed NMR tehnique can be used for estimation of
oil assay in rapeseed. The best results, in respect of precision are obtained using the
conventional Soxhlet extraction. Therefore this method can be useful for the determination
of oil content in rapeseed.

Acknowledgements
This study was supported by University of Gdansk (DS/8270-4-0093-6). The authors
would like to thank Ms Izabela Tomasiak for her help with experimental work and Elstar
Oils S.A. for giving us an opportunity to use NMR analyser.

References
[1] Murphy D. J., TIBTECH, 14 (1996) 206
[2] Taylor S. L., King J. W., List G. R., J. Am. Oil Chem. Soc, 70 (4) (1993) 437
[3] Robertson J. R., Barton F.E. Ibid, 61(1984) 543
[4] Gambhir P. N., Ararwala A. K., Ibid, 62 (1985) 103
[5] PN-EN ISO 542:1997, Oilseeds – Sampling, PKN, Warsaw, 1997
[6] PN-EN ISO 664:1997, Oilseeds – Reduction of laboratory sample to test sample,
PKN, Warsaw 1997
[7] PN-EN ISO 659:1999, Oilseeds – Determination of oil content (Reference method),
PKN, Warsaw 1999
[8] PN-73/R-66164, Determination of the oil contents in the oilseeds and oil seed
extraction meals, PKN, Warsaw 1973
[9] PN-EN ISO 10565:1999, Oilseeds – Simultaneous determination of oil and water
contents – Method using pulsed nuclear magnetic resonance spectrometry, PKN,
Warsaw 1999
[10] Eckschlager K., in: Errors in Chemical Analysis, PWN,Warsaw 1974, 138